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Expression of Membrane Proteins at the Escherichia coli Membrane for Structural Studies

  • Manuela Zoonens
  • Bruno MirouxEmail author
Part of the Methods in Molecular Biology™ book series (MIMB, volume 601)

Abstract

Structural biology of membrane proteins is often limited by the first steps in obtaining sufficient yields of proteins because native sources are seldom. Heterologous systems like bacteria are then commonly employed for membrane protein over-expression. Escherichia coli is the main bacterial host used. However, overproduction of a foreign membrane protein at a non-physiological level is usually toxic for cells or leads to inclusion body formation. Those effects can be reduced by optimizing the cell growth conditions, choosing the suitable bacterial strain and expression vector, and finally co-expressing the target protein and the b-subunit of E. coli adenosine triphosphate (ATP)-synthase, which triggers the proliferation of intracytoplasmic membranes. This chapter is devoted to help the experimenter in choosing the appropriate plasmid/bacterial host combination for optimizing the amount of the target membrane protein produced in its correct folded state.

Key words

Bacterial mutant b-subunit of F-ATPase co-expression internal membrane 

Notes

Acknowledgments

This work was supported by INSERM, CNRS and the “Agence Nationale de la Recherche” (grant 05-JCJC-0092-01 to B.M.).

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Université Paris-7, Laboratoire de Biologie Physico-Chimique des Proteines Membranaires, Institut de Biologie Physico-ChimiqueParisFrance

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